An estuarine neritid gastropod, Clithon corona, a potential reservoir of thermostable direct hemolysin-producing Vibrio parahaemolyticus

An estuarine neritid gastropod, Clithon corona, maintained in UV-irradiated recirculating artificial seawater with a salinity of 15 per mil (%o) was found to retain thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus in the gut at significantly higher levels than TDH-non-producing one for at least 14 days. Another estuarine neritid gastropod, C. sowerbianus, was not able to support the preferential survival of TDH-producing organisms. This evidence suggests that, if TDH-producing vibrios are brought to estuaries inhabited by C. corona, repeated ingestion of V. parahaemolyticus by this gastropod could lead to accumulation of TDH-producing vibrios in the estuaries.     

The dorsal aorta initiates a molecular cascade that instructs sympatho-adrenal specification

The autonomic nervous system, which includes the sympathetic neurons and adrenal medulla, originates from the neural crest. Combining avian blood vessel-specific gene manipulation and mouse genetics, we addressed a long-standing question of how neural crest cells (NCCs) generate sympathetic and medullary lineages during embryogenesis. We found that the dorsal aorta acts as a morphogenetic signaling center that coordinates NCC migration and cell lineage segregation. Bone morphogenetic proteins (BMPs) produced by the dorsal aorta are critical for the production of the chemokine stromal cell-derived factor-1 (SDF -1) and Neuregulin 1 in the para-aortic region, which act as chemoattractants for early migration. Later, BMP signaling is directly involved in the sympatho-medullary segregation. This study provides insights into the complex developmental signaling cascade that instructs one of the earliest events of neurovascular interactions guiding embryonic development.     

Conversion of dextran to cycloisomaltooligosaccharides using an enzyme-immobilized porous hollow-fiber membrane

This paper describes a cycloisomaltooligosaccharide glucanotransferase (CITase)-multilayer-immobilized porous hollow-fiber membrane used as an enzyme bioreactor. Dextran, a substrate with an average molecular mass of 43000, is converted into seven- to nine-glucose-membered cycloisomaltooligosaccharides, effective as a preventive for dental caries, aided by convective transport of the substrate to the vicinity of the enzyme through the pores. Epoxy-group-containing graft chains were uniformly appended onto the surface of pores throughout a porous hollow-fiber membrane by radiation-induced graft polymerization. Subsequently, a diethylamino group was introduced, as an anion-exchange moiety, to the graft chains, which caused the chains to expand toward the interior of the pores due to mutual electrostatic repulsion. The expanding graft chain provided multilayer binding sites for CITase. Fifty-five milligrams of adsorbed CITase per gram of membrane is equivalent to the degree of multilayer binding of 5. Finally, 80% of the multilayer-adsorbed CITase was immobilized via enzymatic cross-linking with transglutaminase to prevent the leakage of enzymes. CITase, with a degree of multilayer immobilization of 4, produced the target cycloisomaltooligosaccharides at a conversion yield of 55% in weight at 310 K during permeation by the dextran solution at a space velocity defined as the permeation rate divided by membrane volume of 6 per hour.     

Inhibitory profile of nonapeptide derived from porcine troponin C against angiotensin I-converting enzyme

A novel angiotensin I-converting enzyme (ACE) inhibitory peptide (RMLGQTPTK; 9mer) from porcine skeletal troponin C was investigated for its inhibitory profile. This peptide was noncompetitive and as hydrophobic as the known ACE inhibitory peptides. Aminopeptidase M quickly hydrolyzed 9mer, resulting in production of MLGQTPTK and LGQTPTK with inhibitory activities similar to those of 9mer. The main hydrolysis product of 9mer with carboxypeptidase A and B was RMLGQTPT showing very weak activity. Most products derived from 9mer hydrolysis by ACE, aminopeptidase, or carboxypeptidase showed weak but definite ACE inhibitory activities. Thus, 9mer was estimated to be a wholly efficient inhibitor with these fragment peptides.     

Deformation of (Mg,Fe)SiO3 post-perovskite and D' anisotropy

Polycrystalline (Mg(0.9),Fe(0.1))SiO3 post-perovskite was plastically deformed in the diamond anvil cell between 145 and 157 gigapascals. The lattice-preferred orientations obtained in the sample suggest that slip on planes near (100) and (110) dominate plastic deformation under these conditions. Assuming similar behavior at lower mantle conditions, we simulated plastic strains and the contribution of post-perovskite to anisotropy in the D' region at the Earth core-mantle boundary using numerical convection and viscoplastic polycrystal plasticity models. We find a significant depth dependence of the anisotropy that only develops near and beyond the turning point of a downwelling slab. Our calculated anisotropies are strongly dependent on the choice of elastic moduli and remain hard to reconcile with seismic observations.     

Distribution of photoperiod-insensitive alleles Ppd-B1a and Ppd-D1a and their effect on heading time in Japanese wheat cultivars

The genotypes of photoperiod response genes Ppd-B1 and Ppd-D1 in Japanese wheat cultivars were determined by a PCR-based method, and heading times were compared among genotypes. Most of the Japanese wheat cultivars, except those from the Hokkaido region, carried the photoperiod-insensitive allele Ppd-D1a, and heading was accelerated 10.3 days compared with the Ppd-D1b genotype. Early cultivars with Ppd-D1a may have been selected to avoid damage from preharvest rain. In the Hokkaido region, Ppd-D1a frequency was lower and heading date was late regardless of Ppd-D1 genotype, suggesting another genetic mechanism for late heading in Hokkaido cultivars. In this study, only 11 cultivars proved to carry Ppd-B1a, and all of them carried another photoperiod-insensitive allele, Ppd-D1a. The Ppd-B1a/Ppd-D1a genotype headed 6.7 days earlier than the Ppd-B1b/Ppd-D1a genotype, indicating a significant effect of Ppd-B1a in the genetic background with Ppd-D1a. Early-maturity breeding in Japan is believed to be accelerated by the introduction of the Ppd-B1a allele into medium-heading cultivars carrying Ppd-D1a. Pedigree analysis showed that Ppd-B1a in three extra-early commercial cultivars was inherited from ‘Shiroboro 21’ by early-heading Chugoku lines bred at the Chugoku Agriculture Experimental Station.     

Diversification and genetic differentiation of cultivated melon inferred from sequence polymorphism in the chloroplast genome

Molecular analysis encouraged discovery of genetic diversity and relationships of cultivated melon (Cucumis melo L.). We sequenced nine inter- and intra-genic regions of the chloroplast genome, about 5500 bp, using 60 melon accessions and six reference accessions of wild species of Cucumis to show intra-specific variation of the chloroplast genome. Sequence polymorphisms were detected among melon accessions and other Cucumis species, indicating intra-specific diversification of the chloroplast genome. Melon accessions were classified into three subclusters by cytoplasm type and then into 12 subgroups. Geographical origin and seed size also differed between the three subclusters. Subcluster Ia contained small-seed melon from Southern Africa and South and East Asia and subcluster Ib mainly consisted of large-seed melon from northern Africa, Europe and USA. Melon accessions of subcluster Ic were only found in West, Central and Southern Africa. Our results indicated that European melon groups and Asian melon groups diversified independently and shared the same maternal lineage with northern African large-seed melon and Southern African small-seed melon, respectively. Cultivated melon of subcluster Ic may have been domesticated independently in Africa. The presence of 11 cytoplasm types in Africa strongly supported African origin of cultivated melon and indicated the importance of germplasm from Africa.     

Molecular cloning and biochemical characterization of medaka (Oryzias latipes) lysosomal neu4 sialidase

Glycoconjugates are known to be involved in many physiological events in vertebrates. Sialidase is one of the glycosidases, which removes sialic acid from glycoconjugates. In mammals, the properties and physiological functions of sialidases have been investigated, while there is little understanding of fish sialidase. Here, to investigate the significance of fish neu4 sialidase, neu4 gene was cloned from medaka brain mRNA and identified. Sialidase-specific motifs (GPG, YRVP and Asp-Box) were well conserved in the medaka neu4 polypeptide. Optimal pH of medaka neu4 sialidase was 4.6, but its activity was sustained even at neutral and weak alkaline pH. The neu4 considerably cleaved sialic acid from 4-methylumbelliferyl-N-acetyl-α-d-neuraminic acid and sialyllactose, but not from ganglioside and fetuin, which are good substrates for human NEU4. neu4 activity was mostly detected in mitochondria/lysosome fraction after biochemical fractionation, and indirect immunofluorescence assays revealed neu4 localization in lysosome in neu4 overexpressed cells. Next, developmental change in medaka neu4 and other sialidase mRNA levels were estimated by real-time PCR. Each sialidases showed different expression patterns in embryonic development: neu4 was up-regulated at late developmental stage in embryo, and neu3a mRNA level was quite high in 0.5 dpf. On the other hand, neu3b expression was drastically increased after hatching, suggesting that each sialidase may play a different role in embryonic development.